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245	1	0	_aE2-Ub-R74G strategy reveals E2-specific ubiquitin conjugation profiles in live cells
490	0		_aNature Chemical Biology (2025). https://doi.org/10.1038/s41589-024-01809-9
500			_aArtículo
520	3		_aThe E2 ubiquitin (Ub)-conjugating enzyme primarily determines Ub conjugation as Ub-isopeptide (lysine), Ub-oxyester (serine/threonine) or Ub-thioester (cysteine). However, E2-specifc Ub conjugation profles within cells remain elusive. Here we developed the fusion E2-Ub-R74G profling (FUSEP) strategy to access E2-specifc Ub conjugation profles in cells with amino acid resolution. The probe-specifc leucine-glycine-glycine-glycine-modifed Ub remnant enables systematic studies of non-lysine Ub conjugation and provides site-specifc information. Multiple E2 enzymes were found to be involved in non-lysine ubiquitination. Profling with UBE2D3-Ub-R74G probes identifed a post-translational modifcation, tyrosine ubiquitination, in human Cullin-1, a scafold protein for Cullin-RING E3 Ub ligases. This modifcation is distinct from lysine ubiquitination. A single-pass membrane-bound E3 ligase, RNF149, was identifed to pair with UBE2D3 to regulate pyroptosis by ubiquitinating apoptosis-associated speck-like protein ASC. The availability of this toolbox paves the way for uncovering E2-specifc Ub conjugation profles and identifying previously unknown E3 Ub ligases for potential therapeutic applications.
650	1	4	_aUB-R74G STRATEGY REVEALS E2-SPECIFIC UBIQUITIN CONJUGATION PROFILES IN LIVE CELLS
700	1	2	_aShen, S.;Yin, H.
856	4	0	_uhttps://drive.google.com/file/d/1UuvGLdi9QQFjEUBO4h6uhSW2My8qFSa1/view?usp=drive_link _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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